Transcriptome of early embryonic invasion at implantation sites in a murine model.

Successful implantation relies on the interaction between a competent embryo and a receptive endometrium. The aim of the present study was to investigate genes differentially expressed in early invasive embryonic tissue versus decidual tissue in mice. Samples were obtained from the ectoplacental cone, the immediately surrounding deciduas and from deciduas from interimplantation sites. Microarray analysis showed that 817 genes were differentially expressed between extra-embryonic tissue and the surrounding decidua and that 360 genes were differentially expressed between the different deciduas, with a high representation of developmental processes. Genes differentially expressed in the maternal compartment included chemokines, lipoproteins, growth factors and transcription factors, whereas the embryonic invasive tissue expressed genes commonly observed in invasive tumour-like processes. These results provide information about genes involved in early embryonic invasion and the control exerted by the surrounding decidua. This information may be useful to find targets involved in pathologies associated with implantation failure and early pregnancy loss.

Agenesia del cuerpo calloso. Discordancia clínico-radiológica. Análisis tras 15 años de experiencia / Agenesis of the corpus callosum. Clinical-radiological discordance. Analysis after 15 years of experience

Introducción. La agenesia del cuerpo calloso (ACC) supone una desconexión interhemisférica por falta de formación de dicha estructura en el desarrollo embrionario. Prevalencia: 0,3-0,5% en población general y 2,3% en personas con discapacidad. Se asocia a prematuridad y edad materna avanzada. Clínica muy variable.Puede confirmarse con RMN. Nuestro objetivo es determinar: 1) la correlación entre las anomalías del CC en neuroimagen y en la clínica y 2) averiguar factores influyentes en estas diferencias. Material y métodos. Estudio retrospectivo (15 años) de casos de ACC de una Unidad de Rehabilitación Infantil. Los datos epidemiológicos, clínicos e imaginológicos recogidos se confrontaron entre sí y se compararon con la bibliografía. Resultados. El 75% eran mujeres. La edad media de la primera y última revisión, fue 1,8 y 10,7 años, respectivamente. Los motivos de consulta principales fueron alteraciones ortopédicas o neuromotoras. No existían antecedentes obstétricos de interés en el 75%. En la clínica, encontramos: retraso psicomotor (37%), alteración del lenguaje (31%), dificultad para marcha o la manipulación (57%), malformaciones axiales (25%). Las patologías periféricas predominantes se dieron en pies y caderas. La prueba complementaria más frecuentemente solicitada fue la RMN. El 75% asociaba otras malformaciones cerebrales.Tratamiento prescrito: fisioterapia (100%), férulas (75%), silla de ruedas (50%), tratamiento logopédico y/o quirúrgico (25%). Evolución: 87% favorable. Conclusiones. 1) La RMN puede ayudar al diagnóstico inicial y despistaje de malformaciones asociadas, pero no siempre se correlaciona con la clínica y no puede servirnos de medidor pronóstico. 2) La ACC puede ser grave a nivel neurológico, psicológico y motriz, sin embargo, en ocasiones cabe esperar una evolución favorable (AU)

Introduction. The agenesis of the corpus callosum (ACC) implies an interhemispheric disconnection due to the lack of formation of said structure in the embryo development. Prevalence: 0.3-0.5% in the general population and 2.3% on in persons with disability. It is associated to prematurity and older maternal age. The clinical variable varies greatly and can be confirmed with the MRI. Our objective has been to determine 1) the correlation between abnormalities in neuroimaging and in the clinical features and 2) to discover the factors influencing these differences. Material and methods. A retrospective study (15 years) of cases of ACC in a Children's Rehabilitation Unit. The epidemiological, clinical and imaging data collected were compared to each other and with the literature. Results. A total of 75% were women. Average age of the first and last check-up was 1.8 and 10.7 years, respectively. The main reasons of the consultations were orthopedic or neuro-motor disorders. There were no obstetric backgrounds of interest (75%). At the clinical features, we found psychomotor retardation (37%), language disturbance (31%), difficulty to walk or with manual handling (57%), axial malformations (25%). The predominant peripheral neuropathy occurred in feet and hips. The most frequently requested complementary test was an MRI. A total of 75% were associated with other brain malformations. Treatment prescribed: physiotherapy (100%), splints (75%), wheelchair (50%), speech therapy treatment and/or surgery (25%). Evolution: 87% favorable. Conclusions. 1) MRI can help the initial diagnosis and screening of associated malformations, but does not always correlate with the clinical features and cannot be used by us to gauge prognosis. 2) The ACC may be serious on the neurological, psychological and motor level, however sometimes we can expect a favorable outcome (AU)

Endometrial receptivity is affected in women with high circulating progesterone levels at the end of the follicular phase: a functional genomics analysis.

BACKGROUND: Elevated serum progesterone levels at the end of the follicular phase in controlled ovarian stimulation (COS) leads to a poorer ongoing pregnancy rate in IVF cycles due to reduced endometrial receptivity. The objective of this study was to use microarray technology to compare endometrial gene expression profiles at the window of implantation according to the levels of circulating progesterone. METHODS: For this prospective cohort study, microarray data were obtained from endometrial biopsies from 12 young healthy oocyte donors undergoing COS with pituitary suppression by either gonadotrophin-releasing hormone (GnRH) agonists or antagonists, and recombinant FSH. On the day of recombinant chorionic gonadotrophin (rCG) administration, six women had serum progesterone levels (P) >1.5 ng/ml (study group) and six had serum P levels <1.5 ng/ml (control group). Endometrial samples were collected using a Pipelle catheter 7 days after the rCG injection. RESULTS: Using the parametric test, we identified 140 genes significantly dysregulated (64 up- and 76 down-regulated) in the study group endometria compared with the control endometria, regardless of the GnRH analogue employed. These genes are related to cell adhesion, developmental processes, the immune system and others, which are all required for normal endometrial function development. Of the 25 gene targets previously proposed as markers for endometrial receptivity, 13 appeared over-regulated in the study group. CONCLUSIONS: Our results reveal that elevated progesterone levels on the day of rCG administration can induce significant alterations in the gene expression profile of the endometrium.

Endometrial gene expression analysis at the time of embryo implantation in women with unexplained infertility.

Successful embryo implantation depends on the quality of the embryo, as well as on the receptivity of the endometrium. The aim of this study was to investigate the endometrial gene expression profile in women with unexplained infertility in comparison with fertile controls at the time of embryo implantation in order to find potential predictive markers of uterine receptivity and to identify the molecular mechanisms of infertility. High-density oligonucleotide gene arrays, comprising 44 000 gene targets, were used to define the endometrial gene expression profile in infertile (n = 4) and fertile (n = 5) women during the mid-secretory phase (day LH + 7). Microarray results were validated using real-time PCR. Analyses of expression data were carried out using non-parametric methods. Hierarchical clustering and principal component analysis showed a clear distinction in endometrial gene expression between infertile and fertile women. In total we identified 145 significantly (>3-fold change) up-regulated and 115 down-regulated genes in infertile women versus controls. Via Database for Annotation, Visualization and Integrated Discovery functional analysis we detected a substantial number of dysregulated genes in the endometria of infertile women, involved in cellular localization (21.1%) and transport (18.8%) and transporter activity (13.1%) and with major localization in extracellular regions (19.2%). Ingenuity Pathways Analysis of the gene list showed dysregulation of gene pathways involved in leukocyte extravasation signalling, lipid metabolism and detoxification in the endometria of infertile women. In conclusion, endometrial gene expression in women with unexplained infertility at the time of embryo implantation is markedly different from that in fertile women. These results provide new information on genes and pathways that may have functional significance as regards to endometrial receptivity and subsequent embryo implantation.

Microarray analysis in sperm from fertile and infertile men without basic sperm analysis abnormalities reveals a significantly different transcriptome.

Sperm analysis following World Health Organization guidelines is unable to explain the molecular causes of male infertility when basic sperm parameters are within a normal range and women do not present gynecologic pathology. Consequently, there is a need for accurate diagnostic tools in this area, and microarray technology emerges as promising. We present, for the first time, preliminary results of a comparison of sperm mRNA expression profiles between fertile and infertile men with normal semen parameters, discovering profound discrepancies between groups, with potential diagnostic and therapeutic possibilities.

The significance of sperm DNA oxidation in embryo development and reproductive outcome in an oocyte donation program: a new model to study a male infertility prognostic factor.

OBJECTIVE: One byproduct resulting from free radical damage is the DNA hydroxylation also known as DNA oxidation. Our aim with this work was to determine the relevance of sperm DNA oxidation on embryo quality in oocyte donation cycles. DESIGN: We prospectively studied pairs of oocyte donation cycles, i.e., the same oocyte donors, donating to two recipients, where the only difference between the two treatments was the use of a different sperm sample. SETTING: University-affiliated private IVF setting. PATIENT(S): Infertile male partners from couples undergoing oocyte donation cycles (n = 38): 76 semen aliquots analyzed before and after semen processing by swim up. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): We measured sperm DNA oxidation by flow cytometry using the OxiDNA assay and correlated it with embryo quality parameters, implantation, and pregnancy outcome. RESULT(S): A positive correlation was seen between embryo fragmentation and DNA oxidation of capacitated samples at 48 hours and 72 hours after fertilization. However, when we analyzed the differences in the IVF outcome parameters of the couples who shared the oocyte cohort (same donor) with the differences in the OxiDNA values, we observed increased and further relationships with cell embryo division 48 hours after fertilization. A negative association with blastocyst formation was also detected. CONCLUSION(S): Oxidative damage in the DNA is clearly increased in samples with lower sperm motility. An association between early and late embryo quality and sperm DNA oxidation supports the relevance of the hydroxylation of 8-oxoguanine as a biomarker of sperm quality reflecting the free radical damage in human sperm.

Identification, characterization and co-localization of label-retaining cell population in mouse endometrium with typical undifferentiated markers.

BACKGROUND: The endometrium, lining of the uterus, is a highly active organ that is remodelled periodically during the lifespan. Different studies suggest the presence of an adult or progenitor stem cell (PSC) population in this tissue because of its cyclic regenerative capacity. METHODS: In this study, we aim at identifying and localizing the putative PSC population in the murine uterus using the 5-bromo-2'-deoxyuridine (BrdU) labelling method to detect label-retaining cells (LRCs) that cycle slowly. Uteri from BrdU-treated mice were analysed via single and double immunohistochemistry to co-localize them with the markers of undifferentiation already described such as c-KIT and POU5F1 (also known as OCT-4). Finally, we confirmed the presence of the indicated markers at mRNA level. RESULTS: We observed the presence and gradual decrease of LRCs in the endometrium during the lifespan of the mice. In adulthood, the LRC population decreased notably and remained in the lower region of the stroma in the murine endometrium. Some of the endometrial LRCs co-localized with c-KIT and POU5F1. PCR and nested-PCR confirmed the presence of these undifferentiated markers. CONCLUSIONS: We demonstrated that the murine endometrium possesses LRCs with the features of a putative PSC population localized at the lower region of the stroma.

A butterfly vertebra or a wedge fracture?

Two patients, who had injuries to the dorsal spine with vertebral wedging, are described. In both a compression wedge fracture was diagnosed, but a careful study of the anteroposterior radiographs showed that the presumed injury was a butterfly vertebra. This rare congenital anomaly, which is usually without clinical significance, may be mistaken for a wedge fracture unless the anteroposterior view is correctly assessed.